中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (24): 4371-4375.doi: 10.3969/j.issn.1673-8225.2010.24.001

• 骨组织构建 bone tissue construction •    下一篇

转化生长因子β1联合骨形态发生蛋白2诱导骨髓间充质干细胞体外向软骨细胞的分化

张清林，吕惠成，吴一民   

  1. 内蒙古医学院第二附属医院骨科创伤2科，内蒙古自治区呼和浩特市  010030
  • 出版日期:2010-06-11 发布日期:2010-06-11
  • 通讯作者: 吴一民，博士，主任医师，教授，内蒙古医学院第二附属医院骨科创伤2科，内蒙古自治区呼和浩特市 010030
  • 作者简介:张清林★，男，1984年生，山东省济宁市人，内蒙古医学院在读硕士，主要从事骨髓间充质干细胞与骨折愈合的研究。 linzi688@163.com

In vitro differentiation from bone marrow mesenchymal stem cells into chondrocytes induced by transforming growth factor-beta combined with bone morphogenetic protein-2

Zhang Qing-lin, Lü Hui-cheng, Wu Yi-min   

  1. Second Department of Orthopedics, Second Affiliated Hospital of Inner Mongolia Medical College, Huhehaote  010030, Inner Mongolia Autonomous Region, China
  • Online:2010-06-11 Published:2010-06-11
  • Contact: Wu Yi-min, Doctor, Chief physician, Professor, Second Department of Orthopedics, Second Affiliated Hospital of Inner Mongolia Medical College, Huhehaote 010030, Inner Mongolia Autonomous Region, China
  • About author:Zhang Qing-lin★, Studying for master’s degree, Second Department of Orthopedics, Second Affiliated Hospital of Inner Mongolia Medical College, Huhehaote 010030, Inner Mongolia Autonomous Region, China linzi688@163.com

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摘要：

背景：国内外许多学者成功利用转化生长因子β1诱导骨髓间充质干细胞向软骨细胞分化报道较多，而对骨形态发生蛋白2作为诱导因子诱导骨髓间充质干细胞向软骨细胞分化的报道较少。
目的：观察验证转化生长因子β1和骨形态发生蛋白2诱导骨髓间充质干细胞下向软骨细胞表型转化的可能性及相互作用。
方法：取健康Wistar大鼠骨髓，采用贴壁法筛选获得骨髓间充质干细胞。按添加生长因子的不同分为4组：转化生长因子β1+骨形态发生蛋白2组，转化生长因子β1组，骨形态发生蛋白2组，对照组不添加任何生长因子。于诱导14 d后，分别进行阿利新蓝染色和二甲基亚甲蓝比色法定性定量检测糖胺聚糖的分泌表达，免疫组织化学法检测软骨特异性Ⅱ型胶原的表达。
结果与结论：3个加入细胞因子的实验组在阿利新蓝染色和软骨特异性Ⅱ型胶原疫组化法检测中均有不同程度的阳性表达，糖胺聚糖的定量检测中，转化生长因子β1+骨形态发生蛋白2联合应用组的糖胺聚糖含量明显高于其他组(P < 0.05)。结果提示转化生长因子β1、骨形态发生蛋白2联合作用更能促进骨髓间充质干细胞向软骨细胞诱导分化，分泌软骨特异性基质，有可能成为软骨组织工程较理想的种子细胞来源。

关键词: 骨髓间充质干细胞, 软骨细胞, 转化生长因子&beta, 1, 骨形态发生蛋白2, 软骨组织工程

Abstract:

BACKGROUND: Multiple studies have reported that transforming growth factor-β1 (TGF-β1) induced differentiation from bone marrow mesenchymal stem cells (BMSCs) into chondrocytes; however, reports addressing bone morphogenetic protein-2 (BMP-2) inducing the differentiation remain less.
OBJECTIVE: To verify the possibility and interaction of differentiation from BMSCs into chondrocytes induced by TGF-β1 and BMP-2.
METHODS: BMSCs were derived from healthy Wistar rats using adherent methods. The BMSCs were assigned into four groups: TGF-β1+BMP-2, TGF-β1, BMP-2, and control groups. At 14 days after induction, alcian blue staining and dimethyl-methylene blue chromatometry were employed to determine the glycosaminoglycan (GAG) level, and immuno-histochemistry was used to detect the expression of specific type II collagen.
RESULTS AND CONCLUSION: In the three experimental groups, alcian blue staining and type II collagen immunohistochemistry examinations showed that chondyocytes could express glycosaminoglycan (GAG) and type II collagen, and the GAG detection showed that the expression of GAG gene was significantly greater in the TGF-β1+BMP-2 group than in the TGF-β1 group and BMP-2 group (P < 0.05). TGF-β1 combined with BMP-2 was more effective for promoting the differentiation from BMSCs into chondrocytes, which might be used as the better source of the seed cells of cartilage tissue engineering.

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